Establishment of monoclonal antibody against human Apo B-48 and measurement of Apo B-48 in serum by ELISA method

Uchida, Yoshiaki; Kurano, Yoshihiro; Ito, Satoru

doi:10.1002/(sici)1098-2825(1998)12:5<289::aid-jcla7>3.0.co;2-1

Cited by 41 publications

(32 citation statements)

References 11 publications

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“…Liporotein(a) (Lp(a)) was measured by a latex agglutination method (Sekisui Medical, Tokyo, Japan). The concentration of apoB48 was measured with a chemiluminescent enzyme immunoassay (Fujirebio, Tokyo, Japan) 23,24) . LPL mass (Daiichi Pure Chemicals, Tokyo, Japan) were measured by ELISA.…”

Section: Biochemical Analysismentioning

confidence: 99%

Acute Effects of Shortly Pre- Versus Postprandial Aerobic Exercise on Postprandial Lipoprotein Metabolism in Healthy but Sedentary Young Women

Hashimoto

Ootani

Hayashi

et al. 2011

JAT

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Section: Biochemical Analysismentioning

confidence: 99%

Acute Effects of Shortly Pre- Versus Postprandial Aerobic Exercise on Postprandial Lipoprotein Metabolism in Healthy but Sedentary Young Women

Hashimoto

Ootani

Hayashi

et al. 2011

JAT

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“…This evidence is supportive of our claim that this ELISA system can measure rabbit apoB-48. It has been reported that treatment of serum with TritonX-100 increases the recognition of apoB-48 epitope by hexapeptide antibodies [2,4,16]. In our previous study, we found that our monoclonal antibody (4C8) showed different characteristics in the recognition site of apoB-48 from the other hexapeptide polyclonal or monoclonal antibodies hitherto reported [10,12,16].…”

Section: Discussionmentioning

confidence: 59%

“…The apoB-48 Cterminal heptapeptide (consisting of an N-terminal cysteine residue attached to the C-terminal hexapeptide equivalent to residues 2147-2152 of apoB-48) was used as the immunogen to raise the polyclonal antibody used in this assay. Uchida et al [16] later developed an apoB-48 ELISA method using the same C-terminal peptide as the immunogen for monoclonal antibody production. No cross-reactivity was found with apoB-100, as verified by ELISA and western blot analyses using polyclonal and monoclonal antibodies.…”

Section: Introductionmentioning

confidence: 99%

Determination of Immuno-Reactive Rabbit Apolipoprotein B-48 in Serum by ELISA

et al. 2010

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is produced by the small intestine, is associated with chylomicrons, and appears to be a suitable marker for clinical studies of postprandial dynamics of lipoproteins. It is also associated with cardiovascular risk factors. We have developed an assay system to quantify immuno-reactive apoB-48 in rabbit serum or plasma. A microtiter-plate was coated with monoclonal antibody raised against human apoB-48 C-terminal specific decapeptide that has high homology to the rabbit C-terminal sequence. Appropriate ELISA standard curves were obtained using apoB-48 extracted from rabbit serum by immuno-affinity chromatography. No cross-reactivity was found with apoB-100 in western blot analyses. Intra-and inter-assay CVs were less than 3%. Recovery of rabbit apoB-48 spiked in serum was within 93.4-105%. ApoB-48 levels in healthy controls rabbits fed a normal diet were within the range of 0.903-1.09 µg/ml (mean ± SD: 1.03 ± 0.084 µg/ml). In healthy animals, the blood apoB-48 level was markedly increased by a high fat diet and in the postprandial state in parallel with serum triglyceride. Ezetimibe, cholesterol absorption inhibitor, given orally to rabbits fed on a high fat diet blocked further increase of blood levels of apoB-48 and triglyceride. This method for measuring apoB-48 using the monoclonal antibody is simple, reliable, and suitable for routine analyses.

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“…A synthetic peptide corresponding to the 10 amino acids of apoB48 C-terminal was used as the antigen for this monoclonal antibody. Uchida et al also created a monoclonal antibody using the 8 C-terminal amino acids by the similar method 24) , and ELISA to measure apoB48 was also reported 25) . Our anti-apoB48 monoclonal antibody (4C8) thus obtained does not recognize apoB100 despite its similarity to apoB48.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of Apolipoprotein B48-Containing Lipoproteins with a Monoclonal Antibody Against Apolipoprotein B48

Yoshimura

Kinoshita

Teramoto

2009

JAT

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Aim: Remnant lipoproteins are well known to play a pivotal role in atherosclerosis. In patients with postprandial dyslipidemia, metabolic pathways for exogenous lipoproteins are generally disturbed, resulting in accumulation of chylomicron remnants. Although it has been difficult to make a specific antibody against apolipoprotein B48 (apoB48), a constituent of exogenous lipoproteins, we succeeded in creating a specific monoclonal antibody against apoB48. In this study, we isolated apoB48-containing lipoproteins from lipoproteins with a density less than 1.019 g/mL using this anti-apoB48 monoclonal antibody (4C8). Methods: Apolipoproteins and lipids were analyzed to confirm whether apoB48-containing lipoproteins are isolated from other lipoproteins. The characteristics of apoB48-containing lipoproteins in the plasma of patients were compared with type 2 diabetes mellitus and non-diabetic patients. Furthermore, the uptake of apoB48-containing lipoproteins by THP-1 cells (a human acute monocytic leukemia cell line), HepG2 cells (a human hepatoma cell line), and human umbilical vein endothelial cells (HUVEC) was investigated. Also, the expression of apoB48 receptors in these cells was tested with RT-PCR. Results: Apolipoprotein analysis of 4C8-bound lipoproteins indicated the isolation of apoB48-containing lipoproteins, because the content of apoB100 was quite low (less than 5%). Compared with lipoproteins that were not bound to the antibody, apoB48-containing lipoproteins had a higher content of triglycerides. There was no significant difference in the composition of apoB48-containing lipoproteins between patients with and without type 2 diabetes. Uptake of fluorescence-labeled apoB48-containing lipoproteins by THP-1-derived macrophages and HepG2 cells, but not by HUVEC, was observed. The specificity of this uptake was confirmed because the fluorescent signal was competed out by an excess amount of the same unlabeled lipoproteins. RT-PCR revealed the expression of apoB48 receptors in THP-1 and HepG2 cells but not in HUVEC. These results suggest that specific uptake of apoB48-containing lipoproteins may occur via apoB48 receptors. Conclusion

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Establishment of monoclonal antibody against human Apo B-48 and measurement of Apo B-48 in serum by ELISA method

Cited by 41 publications

References 11 publications

Acute Effects of Shortly Pre- Versus Postprandial Aerobic Exercise on Postprandial Lipoprotein Metabolism in Healthy but Sedentary Young Women

Acute Effects of Shortly Pre- Versus Postprandial Aerobic Exercise on Postprandial Lipoprotein Metabolism in Healthy but Sedentary Young Women

Determination of Immuno-Reactive Rabbit Apolipoprotein B-48 in Serum by ELISA

Isolation and Characterization of Apolipoprotein B48-Containing Lipoproteins with a Monoclonal Antibody Against Apolipoprotein B48

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